| Combination of ELISA and dried blood spot technique for the quantification of large molecules using exenatide as a model. | |
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MedLine Citation:
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PMID: 21515389 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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INTRODUCTION: To explore the feasibility of coupling dried blood spot (DBS) technique with ELISA for the quantification of large molecules, exenatide was used as a model. A method for the quantification of exenatide in human blood was developed and evaluated. METHODS: Exenatide standard and quality control samples prepared in fresh human blood were spotted on DBS cards and then extracted. The extraction conditions were optimized by comparing different extraction solutions, with/without protease inhibitors, and various incubation times. A competitive ELISA assay was used for quantification of exenatide from DBS samples. RESULTS: The assay range of exenatide standards in blood was 100-5000pg/mL. The intra-assay precision (%CV) was from 1.2% to 16.3%, and the accuracy (%Recovery) was from 87.5% to 117.0%. The inter assay precision (%CV) was from 1.7% to 14.3%, and the accuracy was from 95.0% to 115.5%. All the above assay parameters met acceptance criteria. Furthermore, the storage stability of exenatide on DBS cards was tested at ambient temperature as well as at 4°C and -70°C, and it was found that change of storage temperature did not affect the stability of exenatide significantly. DISCUSSION: Our results demonstrated a successful coupling of DBS technique with ELISA for quantification of exenatide in human blood, and the DBS-ELISA combination has a great potential to be further applied for the quantification of other large molecule drugs or biomarkers. |
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Authors:
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Yong-Qing Lin; Risha Khetarpal; Yilu Zhang; Hengchang Song; Shawn S Li |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2011-4-16 |
Journal Detail:
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Title: Journal of pharmacological and toxicological methods Volume: - ISSN: 1873-488X ISO Abbreviation: - Publication Date: 2011 Apr |
Date Detail:
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Created Date: 2011-4-25 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9206091 Medline TA: J Pharmacol Toxicol Methods Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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Copyright © 2011. Published by Elsevier Inc. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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